Bovine viral diarrhea virus type 1b vaccine compositions and methods
A technology for bovine viral diarrhea and composition, applied in the field of improvement, and/or management of microbial or viral infection or its symptoms, prevention, treatment, and immunogenic composition, capable of solving problems such as inapplicability
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Embodiment 1
[0100] Example 1 - Production of BVDV Modified Live Viruses
[0101] This example provides an illustrative method for large-scale production of BVDV-1b modified live virus (MLV).
[0102] Materials and Methods
[0103] Viruses and Microbes
[0104] From the United States Department of Agriculture (USDA), Animal and Plant Health Inspection Service (APHIS), Center for Veterinary Biologics Laboratory (CVB-L) ( Ames, IA, USA) to obtain the TGAC strain of BVDV-1b. The Master Seed (MS) is subsequently denoted "TVL-BVDV1b MS 04 / 27 / 2007 DW3-095" and, as noted in Applicant's pending U.S. Provisional Patent Application No. 61 / 427,404 filed herewith, has been Deposited under conditions ensuring that persons authorized under 37 C.F.R. § 1.14 and 35 U.S.C. § 122, as determined by the Commissioner of the Patent and Trademark Office, will have access to the culture during the pendency of this patent application. This deposit is available, as required by foreign patent laws, in the countr...
Embodiment 2
[0124] Example 2 - BVDV-1b Monovalent Vaccine Calf Challenge Study
[0125] This example demonstrates the efficacy of the BVDV-lb monovalent vaccine in preventing infection in vaccinated animals.
[0126] Materials and Methods
[0127] animal
[0128] Healthy 4- to 5-month-old mixed-breed heifers were purchased from commercial sources, identified by randomly drawn ear tags, and serologically screened for susceptibility to BVDV. All calves received one dose of ceftiofur crystalline free acid (Excede®, Pfizer Animal Health, New York, NY, USA) upon arrival. Calves had ad libitum access to water, hay and pelleted rations.
[0129] vaccine
[0130] The BVDV-1b vaccine, modified live virus, was prepared as described above. Briefly, BVDV-1b master seed virus was propagated in the TVL-BK cell line (passage 20), harvested at passage 10, stabilized according to the production protocol, filled into vials, and lyophilized. The BVDV concentration was 8 x 10 as determined by the Spear...
Embodiment 3
[0232] Example 3 - BVDV-1b virus backpassage (Backpassage) evaluation
[0233] This example evaluates the stability of TVL-BVDV-1b MSV to assure that it does not restore virulence when administered to animals.
[0234] 1 ampoule of virus was thawed and used to inoculate TVL-BK cells to produce TVL-BVDV-1b+1 (5 × 10 7 TCID 50 / mL). The virus is used in a 1 × 10 6 TCID 50 / ratio per calf The first group of 10 colostrum deficient cows were inoculated intranasally. Calves in groups 2 to 5 were inoculated intranasally with pooled nasal secretions from the previous group of calves.
[0235] Group 1 consisted of 10 calves, and the number of calves in Groups 2 to 5 ranged from 2 to 5 calves, depending on the number of calves shedding virus from Group 1 10 calves. Each calf was maintained in a separate pen, and the primary outcome of the study was the lack of clinical signs of BVDV in calves and the consistent lack of genetic stability of MS over at least five consecutive in vi...
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